System and method for monitoring cardiomyocyte beating, viability, morphology and electrophysiological properties

Abstract

Devices, systems and methods for monitoring excitable cells, such as cardiomyocytes, on microelectrode arrays that couple the electro-stimulation of excitable cells to induce or regulate cardiomyocyte beating and the simultaneous measurement of impedance and extracellular recording to assess changes in cardiomyocyte beating, viability, morphology or electrophysical properties in response to a plurality of treatments.

What is claimed is: 
     
       1. A system for monitoring excitable cells, the system comprising a device with at least one well, each well having a bottom with a nonconductive substrate; a power source configured to deliver an electrical signal capable of electro-stimulating excitable cells; an impedance analyzer; and an extracellular recording amplifier; wherein each well further includes:
 a pair of electro-stimulation electrodes configured to receive the electrical signal from the power source thereby delivering an electro-stimulating signal to the well for electro-stimulation of excitable cells; 
 a pair of interdigitated cell-substrate impedance monitoring electrodes communicatively coupled to the impedance analyzer, thereby permitting cell-substrate impedance monitoring of excitable cells attached to the substrate; and 
 an extracellular recording electrode pair communicatively coupled to the extracellular recording amplifier, thereby permitting extracellular recording of excitable cells attached to the substrate, the extracellular recording electrode pair comprising a circular recording electrode and a unitary one-piece reference electrode, the interdigitated cell-substrate impedance monitoring electrodes positioned between the extracellular recording electrode pair. 
 
     
     
       2. The system according to  claim 1 , wherein a percentage of a surface area of the bottom of the at least one well occupied by the pair of electro-stimulation electrodes is selected from the group consisting of 20% or more, 30% or more, 50% or more, and 70% or more. 
     
     
       3. The system according to  claim 1 , wherein at least one electrode of the pair of electro-stimulation electrodes is also at least one electrode of the extracellular recording electrode pair, thereby permitting electro-stimulation of excitable cells and extracellular recording of cells using a same electrode at different time points. 
     
     
       4. The system according to  claim 1 , wherein the impedance analyzer monitors cell-substrate impedance at 10 millisecond time resolution. 
     
     
       5. The system according to  claim 1 , wherein at least one electrode of the pair of electro-stimulation electrodes is also at least one electrode of the pair of impedance monitoring electrodes, thereby permitting electro-stimulation of excitable cells and cell-substrate impedance monitoring of cells using a same electrode at different time points. 
     
     
       6. The system according to  claim 5 , wherein each electrode structure has a plurality of electrode elements. 
     
     
       7. A method for monitoring excitable cells, the method comprising:
 a) providing the device of  claim 1 ; 
 b) adding a sample of excitable cells that lacks biological tissue to the device; 
 c) electro-stimulating the excitable cells; and 
 d) monitoring cell-substrate impedance of the electro-stimulated cells at 10 millisecond resolution. 
 
     
     
       8. The method according to  claim 7 , wherein the excitable cells comprise cardiomyocytes or cardiomyocyte precursor cells, and cell substrate impedance is monitored to assess cell beating. 
     
     
       9. The method according to  claim 7 , wherein the step of electro-stimulating the excitable cells is performed by pulsing the cells with 1V to 2.5V for 0.5-2 milliseconds. 
     
     
       10. The method according to  claim 7 , wherein cell-substrate impedance is monitored between at least two different electro-stimulation intervals. 
     
     
       11. The method according to  claim 7 , further comprising adding a compound suspected of affecting excitation contraction coupling of the excitable cells to the at least one well and monitoring cells to assess an effect on cell beating. 
     
     
       12. The method according to  claim 7 , wherein the device further comprises an extracellular recording electrode pair of electrodes, the method further comprising extracellular recording of the electro-stimulated cells. 
     
     
       13. The method according to  claim 12 , further comprising adding a compound suspected of affecting excitation contraction coupling of the excitable cells to the at least one well and performing extracellular recording to assess an effect of the compound on cell beating. 
     
     
       14. The method according to  claim 12 , wherein at least one electrode is shared between two pairs of electrodes selected from the group consisting of the pair of electro-stimulation electrodes and the pair of impedance monitoring electrodes, the pair of electro-stimulation electrodes and the extracellular recording electrode pair, and the pair of impedance monitoring electrodes and the extracellular recording electrode pair, further wherein the sharing of the at least one electrode is performed by switching communication to the at least one electrode, further wherein the step of monitoring electro-stimulated cells includes monitoring cell-substrate impedance and extracellular recording of cells. 
     
     
       15. The method according to  claim 14 , further comprising adding a compound suspected of affecting excitation contraction coupling of the excitable cells to the at least one well and monitoring cell-substrate impedance and extracellular recording of cells to assess cell beating.